左旋氨氯地平片在Beagle犬体内的药动学研究

目的建立一种快速、灵敏测定Beagle犬血浆中左旋氨氯地平浓度的高效液相色谱-串联质谱(LC-MS/MS)法,研究左旋氨氯地平片在Beagle犬体内的药动学。方法色谱条件采用Phenomenex Synergi Hydro-RP C18色谱柱(30 mm×2mm,4μm);流动相:0.1%甲酸水–0.1%甲酸甲醇,梯度洗脱;体积流量:0.6 m L/min;柱温:室温;进样量:20μL。质谱条件电喷雾离子源(ESI);多级反应监测(MRM);正离子模式;喷雾气温度(TEM):400℃;雾化气(GS1):344.95k Pa;加热辅助气(GS2):413.7 k Pa;气帘气(CUR):206.85 k Pa;碰撞气(CAD):68.95 k Pa;离子喷雾电压(IS):5 500V;扫描时间:200 ms;用于定量分析的MRM离子对分别为左旋氨氯地平m/z 409.0→238.1,内标硝苯地平m/z 347.0→315.2、347.0→271.3。6只Beagle犬灌服6.8 mg/kg苯磺酸左旋氨氯地平片后,以硝苯地平为内标,血浆样品经醋酸乙酯萃取。绘制左旋氨氯地平的药–时曲线,计算主要药动学参数。结果左旋氨氯地平在0.05~20.00 ng/m L线性关系良好,定量下限为0.05 ng/m L。批内、批间精密度RSD值均小于10%,平均提取回收率为89.3%~93.6%,基质效应为99.9%~102.7%。主要药动学参数Cmax=(6.47±0.72)μg/L,tmax=(2.3±0.5)h,t1/2=(11.0±4.6)h,MRT=(15.6±6.8)h,AUC0-t=(68.81±19.29)h·μg/L,AUC0-∞=(71.58±20.35)h·μg/L。结论本法特异、灵敏、准确、可靠,可用于Beagle犬血浆中左旋氨氯地平的药物浓度测定及左旋氨氯地平片药动力学研究。     

黄体酮缓释制剂在Beagle犬中的血药浓度测定方法的建立及其药代动力学研究

本研究建立了高效液相色谱串联质谱法(LC-MS/MS),测定Beagle犬体内黄体酮的血药浓度,并进行黄体酮缓释微球在Beagle犬体内药代动力学的研究。以醋酸甲地孕酮为内标,蛋白沉淀法处理血样。采用电喷雾离子化源(ESI)以多反应监测(MRM)方式进行正离子检测,监测离子对分别为黄体酮m/z 315.2→97.0,内标醋酸甲地孕酮m/z 385.2→267.1。雄性Beagle犬肌注黄体酮缓释微球,于给药后不同时间点采集静脉血,结合WinNonlin 8.1软件计算相关药代动力学参数。在0.1～500.0 ng·mL^-1内,黄体酮的线性关系良好(r~2=0.998 9);验证样品的批内、批间精密度(RSD)小于13.25%,准确度(RE)在8.92%以内;样品在室温放置12 h、-20℃反复冻融3次、-20℃放置60 d条件下均稳定;提取回收率为71.43%～77.97%。Beagle犬肌肉注射黄体酮缓释微球后, t_max为19.00±25.36 h,C_max为137.72±11.59 ng·mL^-1     

盐酸坦洛新缓释胶囊在犬体内的药动学

目的建立犬血浆中坦洛新浓度的测定方法,并应用于盐酸坦洛新缓释胶囊中坦洛新犬体内的药动学研究。方法液相色谱-串联质谱法(LC-MS/MS)测定犬血浆中的坦洛新浓度。坦洛新血浆样品经乙酸乙酯萃取,Agilent ZORBAX SB-C18柱(150 mm×2.1 mm,5μm)分离,阿立哌唑为内标,流动相A为体积分数为0.1%的甲酸水溶液,B为乙腈,线性梯度洗脱,电喷雾电离源(ESI),以多反应离子监测(multiple reaction monitoring,MRM)方式进行正离子检测,用于分析的定量离子分别为m/z409→m/z228(坦洛新),m/z 447.5→m/z 284.8(内标:阿立哌唑)。结果犬血浆中坦洛新的线性为0.1～20.0μg.L-1,定量下限为0.1μg.L-1,日内和日间精密度(RSD)均小于13.59%,准确度(relative error,RE)为-2.54%～4.27%。结论本方法适用于盐酸坦洛新缓释胶囊在犬体内的药动学研究。     

HPLC-MS/MS法检测人血浆中游离花生四烯酸浓度

目的:建立测定人血浆中花生四烯酸浓度的液相色谱串联质谱法。方法:以奥美拉唑为内标,待测血浆经乙酸乙酯萃取后,采用Agilent SB-C18(2.1 mm×100 mm,3.5μm)色谱柱,以水-甲醇(10∶90,v/v)为流动相,流速0.2 mL.min-1,柱温25℃;采用电喷雾离子源(ESI)负离子模式将样品离子化,多反应监测(MRM)模式下对花生四烯酸(m/z 303.2→259.2)和奥美拉唑(m/z 344.2→191.1)进行测定。结果:花生四烯酸在497.88～4997.24 ng.mL-1范围内线性关系良好(r=0.9989)。低、中、高浓度的平均回收率均大于70%,批内与批间RSD均小于10%。结论:本方法简单、快速、灵敏,适用于大样本量血浆中游离花生四烯酸的分析研究。     

UPLC-MS/MS法同时检测人血浆中美托洛尔和曲美他嗪浓度

目的:建立同时检测人血浆中美托洛尔和曲美他嗪浓度的超高效液相色谱-串联质谱(UPLC-MS/MS)方法,应用于临床美托洛尔和曲美他嗪血药浓度监测.方法:血浆采用甲醇沉淀蛋白方法处理后进样分析,以苯佐卡因为内标,色谱柱为Agilent Poroshell 120 EC-C18(100 mm×2.1 mm,2.7 μm),...     

UPLC-MS/MS测定犬血浆中的氢溴酸加兰他敏

目的采用超高效液相色谱-串联质谱法测定犬血浆中氢溴酸加兰他敏(GH)的含量。方法采用BEH C18色谱柱(50 mm×2.1 mm,1.7μm),流动相为乙腈-0.01 mol·L-1醋酸铵(含0.1%甲酸)(40∶60),流速0.1 mL·min-1,柱温40℃,进样量3μL。GH及内标盐酸克仑特罗的监测离子分别为:m/z 288.23→198.06,m/z 277.08→202.93。结果 GH的线性范围为2.38～609.60μg·L-1(r=0.998,n=5),在人血浆基质中,高、中、低浓度(4.76、38.10、304.80μg·L-1)的日内、日间RSD均小于15%,方法的平均回收率为102.3%;血浆基质对测定无干扰。结论所建方法处理简单、快速、灵敏、特异性高,定量准确,适用于血浆中GH的测定。     

HPLC-MS/MS法测定人体血浆中吲哒帕胺的浓度

目的:建立一种简便、灵敏的测定人体血浆中吲哒帕胺浓度的高效液相色谱-串联质谱(HPLC-MS/MS)方法。方法:以瑞格列奈钙为内标,采用乙腈:0.1%甲酸水溶液=90∶10为流动相,以Agilent-ZORBAX-C18柱(2.1mm×150mm,5.0m)色谱柱为分析柱,通过电喷雾离子源(ESI),以正离子多反应监测(MRM)方式进行进样检测。检测离子为[M+H]+m/z 364.1m/z 188.8(吲哒帕胺)和[M+H]+m/z 451.1m/z 134.8(瑞格列奈钙,内标)。结果:吲哒帕胺线性范围为0.5～50ng/mL,定量下限为0.5ng/mL,线性关系良好;高、中、低三个浓度的日内、日间的RSD均<15%,平均回收率为94.60%～110.33%。结论:本方法专属性强,操作简便,灵敏度高,适用于吲哒帕胺制剂的临床药动学研究。     

高效液相串联质谱法测定大鼠血浆中咪哒唑仑的浓度

目的:建立测定大鼠血浆中咪哒唑仑浓度的液相色谱串联质谱法。方法:以伊曲康唑为内标,色谱柱为XTerra MSC18(150mm×2.1mm,3.5μm);流动相为乙腈-0.02%醋酸铵(75∶25);流速为0.2mL.min-1;质谱仪为电喷雾-三重四极杆质谱,以多反应监测(MRM)方式对咪哒唑仑(m/z326.2→m/z291.2)和伊曲康唑(m/z705.3→m/z392.1)进行测定。结果:咪哒唑仑在125～50 000μg.L-1范围内线性关系良好(r=0.996 8),最低检测限为5.0μg.L-1。低、中、高浓度的平均回收率为97.22%,101.88%,90.57%;日内及日间精密度均小于10%。结论:本方法简单、快速、灵敏,能有效检测大鼠血浆中咪哒唑仑的浓度。     

液相色谱-串联质谱法测定Beagle犬血浆中升麻苷H-1的浓度

目的:建立液相色谱-串联质谱的方法测定Beagle犬血浆中升麻苷H-1的浓度。方法:采用20(S)-人参皂苷Rg3为内标,血浆样品经液-液萃取后,以甲醇和水为流动相,梯度洗脱进行分离。通过液相色谱-串联质谱法以多反应监测(MRM)方式检测。用于定量的离子反应分别为m/z615.5→369.6(升麻苷H-1)和783.5→161.1(内标人参皂苷Rg3)。结果:血浆中升麻苷H-1的线性范围为0.5～1000 ng·mL-1,定量下限为0.5 ng·mL-1。在升麻苷H-1浓度为2.0,50,500ng·mL-1时的提取回收率分别为90.4%,94.0%,89.0%。方法的日内和日间精密度(RSD)分别为5.9%,4.7%,3.8%和10.8%,6.3%,7.8%。结论:该方法灵敏、准确,重复性好,可用于Beagle犬血浆中升麻苷H-1的浓度测定。     

液质联用法测定犬血浆中阿普唑仑及其代谢物α-羟基阿普唑仑的浓度

目的建立测定Beagle犬血浆中阿普唑仑及其代谢物α-羟基阿普唑仑浓度的液相色谱-质谱联用(LC-MS)法。方法血浆样品采用1 mol·L~(-1)硼酸盐缓冲液(pH 9.0)碱化、乙酸乙酯-正庚烷(85:15,V:V)萃取后LC-MS测定。色谱柱:Zorbax SB-C_(18)柱(150 mm×3 mm,3.5μm);流动相:乙腈-0.01 mol·L~(-1)乙酸胺缓冲液(含1%甲酸)(45:55,V:V);流速:0.3 mL·min~(-1);柱温:40℃。采用电喷雾正离子模式离子化,用于定量分析的离子分别为m/z 309.2(阿普唑仑)、m/z 325.2(α-羟基阿普唑仑)和m/z 343.2(三唑仑,内标)。结果阿普唑仑和α-羟基阿普唑仑的线性范围分别为0.5～50μg·L~(-1)和0.5～32μg·L~(-1),两者定量下限均为0.5μg·L~(-1),提取回收率均>80%,方法回收率为97.3%～102.5%,批内RSD≤10.4%,批间RSD≤12.2%。结论本方法灵敏、准确、重现性好,适用于阿普唑仑犬体内药动学研究。     

不同保存时间分离的血浆临床疗效研究

目的探讨在(4±2)℃保存条件下,分离出不同保存时间内采用CPDA血袋保存液保存的全血的血浆临床治疗效果.方法将输注悬浮红细胞＞15U的急性大失血患者随机分组:输注8h内分离的新鲜冰冻血浆(FFP)组,输注8～12h内分离的普通冰冻血浆组和输注12h后分离的普通冰冻血浆组.部分血小板低下患者加输新鲜单采血小板或冰冻单采血小板.测定患者输前1h和输后2h指标:①凝血酶时间(TT);②活化部分凝血活酶时间(APTT);③凝血酶原时间(PT);④纤维蛋白原(Fbg).结果输注8～12h内分离的普通冰冻血浆组与输注FFP组比较:TT、PT、APTT及Fbg均无显著性差异(P＞0.05).输注12h后分离的普通冰冻血浆组与输注FFP组比较:TT、PT及Fbg均无显著性差异(P＞0.05),APTT有显著性差异(P＜0.05).结论当FFP不能满足临床需要时,临床上可以采用8～1 2h内分离的普通冰冻血浆替代FFP输注.     

Differences in the binding of drugs to plasma proteins from newborn and adult man. II

Summary The binding of certain drugs to isolated fractions of plasma proteins obtained from newborn and adult man has been studied by equilibrium dialysis. For thiopental, desipramine, nitrofurantoin, sulfamethoxydiazine, meticillin and salicylic acid no difference was found between binding to the albumin fraction from newborns and adults. However, for thiopental, desipramine and promethazine binding to the globulin fraction was smaller in the newborns than in adults. Addition of bilirubin to the albumin fraction decreased the binding of nitrofurantoin, sulfamethoxydiazine and meticillin. No difference in the binding of meticillin to the albumin or globulin fractions from newborns and adults was found. The binding decreased, however, if both fractions were combined. Four mechanisms to explain the difference in binding between newborns and adults are discussed: (1) Displacement of drugs by bilirubin, (2) different binding properties of cord and adult albumin, (3) different properties of the globulins and (4) interaction of albumin with globulins in the newborn.This study was partially supported by grant Ku 156/4 of the Deutsche Forschungsgemeinschaft     

肝素钠的兔血浆与羊血浆效价测定平行性考察

目的：采用兔血浆法和羊血浆法测定肝素钠的效价，比较两种方法在临床操作过程中的可行性并考察两种方法测定结果的平行性。方法：采用兔血浆和羊血浆法。结果：羊血浆法比兔血浆法易于掌握，误差较小。结论：采用羊血浆法优于兔血浆法。     

肝素钠的兔血浆与羊血浆效价测定平行性考察

目的 :采用兔血浆法和羊血浆法测定肝素钠的效价 ,比较两种方法在临床操作过程中的可行性并考察两种方法测定结果的平行性。方法 :采用兔血浆和羊血浆法。结果 :羊血浆法比兔血浆法易于掌握 ,误差较小。结论 :采用羊血浆法优于兔血浆法     

A single LC-MS/MS validated method for tulathromycin quantification in plasma,seminal plasma,and urine to be applied in a pharmacokinetic study in bull

Tulathromycin is a macrolide antibiotic generally used for the treatment of respiratory diseases in cattle and swine. This work proposes an improvement of a previously published LC-MS/MS method for tulathromycin determination in pig serum, here validated in three different bull matrices: plasma, seminal plasma, and urine. The approach is based on a quick protein precipitation with acetonitrile, filtration, and sample dilution before injection, allowing to rapidly process large batches of samples. Analytes separation was obtained using a BEH C18 (50 × 2.1 mm, 1.7 μm) column, maintained at 40°C with a chromatographic run of 5 min. The method was fully validated over concentration ranges suitable for field levels of tulathromycin found in each matrix (0.01–1 μg/ml for plasma, 0.05–5 μg/ml for seminal plasma, and 0.1–10 μg/ml for urine), showing good linearity during each day of testing (R² always >0.99). Accuracy and precision were within ±15% at all QC concentrations in all the three matrices. Furthermore, the use of tulathromycine-d7 as internal standard mitigated the potential impacts of matrix effect. The validated technique was successfully applied to samples collected during a pharmacokinetic study in bulls, allowing to monitor tulathromycin concentrations over time in the three matrices. To our knowledge, this is the first validated approach for LC-MS/MS quantification of tulathromycin in seminal plasma and urine.     

Effect of diltiazem on plasma glucose,insulin and glucagon during an oral glucose tolerance test in healthy volunteers

Summary The effect of the calcium antagonist, diltiazem, on glycoregulation was investigated in 12 healthy volunteers using a standard glucose tolerance test. No significant change was observed in plasma glucose, insulin or glucagon levels after oral treatment for B days with diltiazem 180 mg/day.     

Standardized mental stress in healthy volunteers induced by delayed auditory feedback (DAF)

Summary Using delayed auditory feedback (delay 0.175 s) a standardized form of mental stress was investigated in 8 healthy male volunteers. After a resting period and a period of undelayed reading, the volunteers were exposed for 5 min to the DAF stress. During the DAF period heart rate increased by 10% and systolic and diastolic blood pressure increased by 9% and 18%, respectively. As a measure of acute sympathetic activation, plasma concentrations of norepinephrine and epinephrine rose by 68% and 49%, respectively. The activity of dopamine--hydroxylase in plasma was increased by 25%. From these results it can be concluded that the DAF procedure provides a suitable method for inducing a standardized mental stress in normal subjects, which can be measured as changes in biochemical and cardiovascular variables.Preliminary results were presented at the 49th Spring Meeting of the Deutsche Physiologische Gesellschaft (Badian et al., 1978)     

Potential cellular targets and antibacterial efficacy of atmospheric pressure non-thermal plasma

Atmospheric pressure non-thermal plasma (APNTP) has been gaining increasing interest as a new alternative antibacterial approach. Although this approach has demonstrated promising antibacterial activity, its exact mechanism of action remains unclear. Mechanistic elucidation of the antimicrobial activity will facilitate development and rational optimisation of this approach for potential medical applications. In this study, the antibacterial efficacy of an in-house-built APNTP jet was evaluated alongside an investigation of the interactions between APNTP and major cellular components in order to identify the potential cellular targets involved in plasma-mediated bacterial destruction mechanisms. The investigated plasma jet exhibited excellent, rapid antibacterial activity against a selected panel of clinically significant bacterial species including Bacillus cereus, meticillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and Pseudomonas aeruginosa, all of which were completely inactivated within 2 min of plasma exposure. Plasma-mediated damaging effects were observed, to varying degrees, on all of the investigated cellular components including DNA, a model protein enzyme, and lipid membrane integrity and permeability. The antibacterial efficacy of APNTP appears to involve a multiple-target mechanism, which potentially reduces the likelihood of emergence of microbial resistance towards this promising antimicrobial approach. However, cellular membrane damage and resulting permeability perturbation was found to be the most likely rate-determining step in this mechanism.     

改良RP-HPLC法测定血浆中罗哌卡因的含量

目的:采用反相高效液相色谱法测定犬血浆中罗哌卡因的浓度。方法:待测样品经蛋白沉淀处理后,外标法进行高效液相紫外检测,选用乙腈作为蛋白沉淀剂,色谱柱采用Intertsil C18柱(4.6 mm×250 mm,5μm),流动相为甲醇-水相,体积比为50 50,水相为0.005 nmol.ml-1己烷基磺酸钠,冰醋酸调节pH至3.5,检测波长为215 nm,流速为:1.4ml.min-1;柱温为室温;柱压为2 800 PSI,进样体积为10μl,灵敏度为0.02AUFS。结果:在选定的色谱条件下,罗哌卡因与血浆中的杂质分离良好,罗哌卡因的保留时间为9.8 min。罗哌卡因的检测浓度线性范围为0.1~25μg.ml-1(r=0.9992),最低检测浓度为0.05μg.ml-1,回归方程为Y=16212.66X+37016.53,相关系数r=0.9992,回收率为91.2%~93.6%,RSD为2.10%~3.40%。结论:本法用于罗哌卡因类药物给药后血药浓度的检测,操作简便、可靠、快捷、准确,重现性好。     

Sequential changes in plasma renin activity and plasma catecholamines in mildly hypertensive patients during acute,furosemide-induced body-fluid loss

Summary To evaluate the role of adrenergic mechanisms in the acute response of renin to furosemide, plasma renin activity (PRA) and plasma catecholamine concentrations were measured for 3 h after i.v. administration of furosemide 1 mg/kg to 8 patients with mild essential hypertension. Furosemide induced a prompt and long-lasting increase in renin, with PRA more than doubled at all times. The increase in PRA within the first 30 min paralleled the peak increases in urinary water and sodium flow rates, and significant decreases in plasma volume and central venous pressure. There was no change in plasma catecholamine concentrations. Plasma noradrenaline was increased significantly at 60 min and adrenaline at 90 min, once furosemide had induced a marked loss of body-fluid and 65% decrease in central venous pressure. Both catecholamines remained elevated until the end of the study, whereas urinary water and sodium flow rates had returned to their pre-treatment values by 150 min. Mean blood pressure was essentially unchanged throughout the study, whereas heart rate increased significantly after 90 min. The findings suggest that in mildly hypertensive patients adrenergic mechanisms are not involved in the initial renin response to furosemide, but they come into play later, probably as a result of reflex sympathetic activation triggered by marked volume depletion.